Linked Life Data

10662626

Source:http://linkedlifedata.com/resource/pubmed/id/10662626

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
2
pubmed:dateCreated
2000-3-8
pubmed:abstractText
We present a system for creating influenza virus by generating viral RNA (vRNA) and mRNA from one template. Recently, a system for the generation of influenza A virus entirely from cloned cDNAs was established (Neumann et al., 1999, Proc. Natl. Acad. Sci. USA 96, 9345-9350). Cells were transfected with plasmids for RNA polymerase I-driven intracellular synthesis of all eight viral RNAs, and with protein expression plasmids for the synthesis of viral structural proteins. Although this system is highly efficient in virus generation, the construction and cotransfection of 17 plasmids is cumbersome and may limit the use of this system to cell lines that can be transfected with high efficiencies. Synthesizing both vRNA and mRNA from one template would reduce the number of plasmids required for virus generation. Therefore, we generated a bidirectional transcription construct that contains cDNA encoding PB1 flanked by an RNA polymerase I (pol I) promoter for vRNA synthesis and an RNA polymerase II (pol II) promoter for mRNA synthesis. The utility of this approach is proved by the generation of virus after transfecting the pol I/pol II-promoter-PB1 construct together with vRNA- and protein-expression constructs for the remaining seven segments. Because this approach reduces the number of plasmids required for virus generation, it also reduces the work necessary for cloning, probably enhances the efficiency of virus generation, and expands the use of the reverse-genetics system to cell lines for which efficient cotransfection of 17 plasmids cannot be achieved.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Feb
pubmed:issn
0042-6822
pubmed:author
pubmed:copyrightInfo
Copyright 2000 Academic Press.
pubmed:issnType
Print
pubmed:day
15
pubmed:volume
267
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
310-7
pubmed:dateRevised
2008-11-21
pubmed:meshHeading
pubmed-meshheading:10662626-Animals, pubmed-meshheading:10662626-Cell Line, pubmed-meshheading:10662626-DNA, Recombinant, pubmed-meshheading:10662626-Eukaryotic Cells, pubmed-meshheading:10662626-Gene Expression Regulation, Viral, pubmed-meshheading:10662626-Genetic Vectors, pubmed-meshheading:10662626-Genome, Viral, pubmed-meshheading:10662626-Humans, pubmed-meshheading:10662626-Orthomyxoviridae, pubmed-meshheading:10662626-Plasmids, pubmed-meshheading:10662626-Promoter Regions, Genetic, pubmed-meshheading:10662626-RNA, Messenger, pubmed-meshheading:10662626-RNA, Viral, pubmed-meshheading:10662626-RNA Polymerase I, pubmed-meshheading:10662626-RNA Polymerase II, pubmed-meshheading:10662626-Reverse Transcriptase Polymerase Chain Reaction, pubmed-meshheading:10662626-Templates, Genetic, pubmed-meshheading:10662626-Transcription, Genetic, pubmed-meshheading:10662626-Viral Proteins, pubmed-meshheading:10662626-Virus Replication
pubmed:year
2000
pubmed:articleTitle
"Ambisense" approach for the generation of influenza A virus: vRNA and mRNA synthesis from one template.
pubmed:affiliation
Department of Virology and Molecular Biology, St. Jude Children's Research Hospital, 332 North Lauderdale, Memphis, Tennessee 38105-2794, USA.
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, P.H.S., Research Support, U.S. Gov't, Non-P.H.S., Research Support, Non-U.S. Gov't